The American Civil Liberties Union of Ohio and the Freedom from Religion Foundation filed a lawsuit Thursday in U.S. District Court in Columbus on behalf of an unidentified student in Jackson and his parents, The Columbus Dispatch reported.
The Jackson Board of Education rejected a recent request to remove the painting, which has hung in the Jackson Middle School for almost 70 years.
"The school system was warned weeks ago that this religious display is an unconstitutional endorsement of religion and must be removed," said James Hardiman, the ACLU's legal director. "They have chosen to continue displaying the portrait, making legal action necessary."
Superintendent Phil Howard said Liberty Foundation, a group that defends displays of religious belief, has been working with the district to determine whether the picture can remain. He said he was "shocked" by the filing of the lawsuit.
About 300 people attended the board meeting where the painting was discussed, with most of them opposed to removing it.
Campus cop fatally shoots Texas student during traffic stop
Exploding whale video goes viral on Internet